JP6893413B2 - Lactic acid bacteria-containing fat and oil composition and its manufacturing method - Google Patents

Description

The present invention relates to a lactic acid bacterium-containing oil / fat composition containing a living lactic acid bacterium and a method for producing the same.

Lactic acid bacteria are bacteria that produce lactic acid from sugar. Bacteria that ferment sugar with lactic acid include genus Lactobacillus, Enterococcus, Lactococcus, Pediococcus, Leuconostoc and the like. In addition, the genus Bifidobacterium is known as a bacterium that decomposes sugar to produce lactic acid and acetic acid. Bifidobacterium bacteria may be classified as bifidobacteria separately from lactic acid bacteria, but in the present specification, bifidobacteria bacteria are also included in lactic acid bacteria.
Various microorganisms inhabit the human intestine, and lactic acid bacteria of the genus Lactobacillus and Bifidobacterium are detected in almost all human intestines.

Probiotics are defined by the British microbiologist Fuller as "living microbes that have beneficial effects on humans by improving the balance of the intestinal flora" and reach the intestines alive and multiply in the intestinal environment. It is preferable that it is possible. Lactic acid bacteria are attracting attention as probiotics because they regulate the intestinal flora and also regulate the immune system, endocrine system, nervous system, and so on.
It is desirable to ingest lactic acid bacteria for health promotion and maintenance, and yogurt, lactic acid bacteria beverages and the like are generally used as foods containing lactic acid bacteria. In addition to these, lactic acid bacteria-containing foods as probiotics that are easier and more palatable meet the needs of modern people who demand all of health, convenience, and palatability.
Patent Document 1 discloses chocolate containing lactic acid bacteria including live bifidobacteria, but discloses a technique for improving stability from the time of production, and discloses a survival rate of lactic acid bacteria in the production process and intake. It does not improve the survival rate of lactic acid bacteria in later digestion and absorption.

Lactic acid bacteria are generally damaged by gastric acid when ingested by mouth, and the number of bacteria that reach the intestines alive is drastically reduced compared to when ingested. Therefore, in the case of pharmaceuticals, by setting them to be taken after meals or processing them into enteric-soluble preparations, they are devised so that they reach the intestines alive, and the effect as probiotics is enhanced. However, in foods, it is not possible to set a dosage like a pharmaceutical product, and more palatability is required.

Patent Document 2 discloses a method for producing a stable viable bacterial preparation, which comprises suspending a dry powder containing viable bacteria having an intestinal regulating action in an oil or fat. However, the obtained viable bacterial preparation needs to be used by packing it in a bottle, tube or the like, or by filling it in a soft capsule, and the number of viable bacteria is greatly reduced after long-term storage at room temperature. Further, Patent Document 3 describes a lactic acid bacterium tablet obtained by mixing live lactic acid bacterium powder and a specific type of additive and coating the tablet with an enteric substance such as hydroxymethyl cellulose. In Patent Document 5, lactic acid bacteria cells, gelatin and pectin are described in Patent Document 5, which is an enteric granulated product coated with a layer containing fats and oils and an excipient, a layer containing a hydrous alcohol-soluble protein such as zein, and a layer containing saccharides. An immunostimulatory agent characterized by being encapsulated in an enteric-coated capsule in which alginic acid, cellulose, etc. are combined is described in Patent Document 6 as a seed containing lactic acid bacteria with a water-mixable coating base such as sodium alginate. Although enteric coated granules produced by coating are disclosed, the manufacturing process is complicated and lacks in convenience. Patent Document 7 discloses an enteric-coated lactic acid bacterium composition containing lactic acid bacterium cell powder and polyglycerin fatty acid ester, but as a result of a test using an artificial digestive juice, the number of lactic acid bacterium viable bacteria is low. The result is.

Among the lactic acid bacteria, those used in the production of dairy products such as yogurt and expected to have a health-promoting effect are Lactobacillus brevis, Lactobacillus acidophilus, Lactobacillus gasseri, and Lactobacillus gasseri. Enterococcus faecalis), Bifidobacterium longum, Leuconostoc mesenteroides, and the like are known.
In the genus Lactobacillus brevis species, Lactobacillus brevis subsp. Coagulans (commonly known as "Labre bacterium") and the like are known. In addition, Lactobacillus brevis species have immunostimulatory action and anti-influenza virus activity, and are said to be effective in suppressing weight gain, suppressing liver fat weight gain, and suppressing blood cholesterol increase. It is a lactic acid bacterium that can be expected to have various health effects. Leuconostoc mesenteroides is a lactic acid bacterium that has an immunostimulatory effect and can be expected to have health effects. Lactobacillus acidophilus, Lactobacillus gasseri, Enterococcus faecalis or Bifidobacterium longum are all used in the production of dairy products such as yogurt and are expected to have health-promoting effects.

Japanese Unexamined Patent Publication No. 08-126473 Japanese Unexamined Patent Publication No. 56-002908 Japanese Unexamined Patent Publication No. 04-014434 Japanese Unexamined Patent Publication No. 05-186335 Japanese Unexamined Patent Publication No. 11-199494 Special Table 2002-505251 Japanese Unexamined Patent Publication No. 2001-064189

Nippon Shokuhin Kagaku Kogaku Kaishi 48 (9) pp. 656-663 (2001)

An object of the present invention is to provide a food and a method for producing the same, which can easily and deliciously ingest a living lactic acid bacterium and contribute to health promotion.

By adding a living lactic acid bacterium to the fat and oil composition, we succeeded in producing an excellent lactic acid bacterium-containing fat and oil composition having high survival rate and acid resistance of the lactic acid bacterium.
In addition, after tempering, by adding lactic acid bacteria powder to the chocolate dough before molding, we succeeded in producing lactic acid bacteria-containing chocolate containing live lactic acid bacteria, and contained lactic acid bacteria containing live lactic acid bacteria. We have provided chocolate.

For the first time in the present invention, we have succeeded in producing a lactic acid bacterium-containing oil / fat composition having an extremely high survival rate of lactic acid bacteria. Surprisingly, the lactic acid bacterium-containing oil / fat composition of the present invention has a higher survival rate of lactic acid bacteria than ingesting lactic acid bacteria with yogurt or the like, and the lactic acid bacteria easily reach the intestine alive. Since lactic acid bacteria are usually killed when they are present in gastric acid for 30 minutes, most of the lactic acid bacteria are killed in the stomach even if yogurt or the like is ingested, but the lactic acid bacteria in the lactic acid bacterium-containing oil / fat composition of the present invention survive in gastric acid. Therefore, when the lactic acid bacterium-containing oil / fat composition of the present invention is ingested, most of the lactic acid bacteria can reach the intestine alive.

Furthermore, it was surprisingly found that the lactic acid bacterium-containing oil / fat composition of the present invention has a very long best-by date of one year at room temperature, and more than 70% of the lactic acid bacteria survive even after one year. .. On the other hand, yogurt and lactic acid bacteria beverages are refrigerated and have a best-by date of about 2 weeks. Therefore, the lactic acid bacterium-containing oil / fat composition of the present invention is advantageous in physical distribution such as storage, transportation, and store display, and is easily preferred throughout the year.

It is a figure explaining the manufacturing method of the lactic acid bacterium-containing chocolate. It is a figure explaining the manufacturing method of the lactic acid bacterium-containing almond chocolate. It is a figure explaining the manufacturing method of the lactic acid bacterium-containing fat composition. It is a figure which shows the result of the artificial gastric juice resistance test of the chocolate containing lactic acid bacteria in all of (a), (b) and (c). It is a figure which shows the test result of the artificial gastric juice resistance of lactic acid bacterium-containing chocolate and lactic acid bacterium powder.

One embodiment of the present invention is a lactic acid bacterium-containing oil / fat composition containing a living lactic acid bacterium. The lactic acid bacterium-containing oil / fat composition ^{preferably contains 1 × 10 4} pieces / gram or more of live lactic acid bacteria, and further contains 1 × 10 ⁴ pieces / gram or more and 1 × 10 ¹² pieces / gram or less. Is desirable, and further, 1 × 10 ⁶ pieces / gram or more and 1 × 10 ¹² pieces / gram or less, further 1 × 10 ⁷ pieces / gram or more and 1 × 10 ¹² pieces / gram or less, and further 4 × 10 ⁷ pieces. / Gram or more 1 × 10 ¹² pieces / gram or less is preferable.
Examples of lactic acid bacteria are Lactobacillus brevis, Lactobacillus casei Shirota, Lactobacillus acidophilus L-92, Lactobacillus cremoris, Lactobacillus herveticas, Lactobacillus salivarius, Lactobacillus gasseri OLL2716, Lactobacillus gasselli PA-3, Lactobacillus gasseri PA-3, Lactobacillus bulgaricus OLL1073R-1, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus crispatas, Lactobacillus eugluti, Lactobacillus delbrucky Subspices Bulgaricus 2038, Lactobacillus delbrucky Lactobacillus spp., Such as Bacillus plantarum, Streptococcus spp., Streptococcus Thermophilus 1131, Bifidobacterium longum BB536, Bifidobacterium longum SBT2928, Bifidobacterium lactis GCL2205, Bifidobacterium breve, Bifidobacterium Bactobacillus Infantis, Bifidobacterium Address Sentis, Bifidobacterium Bifidum, Bifidobacterium Catenulatum, Bifidobacterium Pseudocatenuratam, Bifidobacterium Anglertam, Bifidobacterium Galicum, Bifidobacterium Lactobacillus spp., Lactobacillus lactis, Lactobacillus lactis, Lactobacillus lactis, Lactobacillus lactis, Lactobacillus lactis, Lactobacillus lactis, Lactobacillus lactis, Lactobacillus lactis, Lactobacillus lactis, Lactobacillus lactis, Lactobacillus lactobacillus. , Such as Lactobacillus spp., Pediococcus pentosaseus, Pediococcus damnosus, etc. Pediococcus spp. Nostock spp., Etc. can be mentioned.
Among them, the lactic acid bacteria in the present invention are preferably Lactobacillus, Bifidobacterium, Enterococcus, and Leukonostock, and more preferably Lactobacillus brevis, Lactobacillus acidophilus, Lactobacillus gasseri, Enterococcus faecalis, and bi. Bifidobacterium longum and leuconostock mecenteroides, but are not limited to bacteria that produce lactic acid.
The fat and oil composition in the present invention refers to a composition mainly composed of fats and oils, and contains 20 to 99%, more preferably 30 to 60% of fats and oils in the entire composition. In addition to fats and oils, fats and oils compositions are produced by using sugars, cacao-derived raw materials, milk-derived raw materials, flavors, emulsifiers, etc. as raw materials as necessary, and crushing, mixing, solidifying, etc., as necessary. Will be done.
Examples of the lactic acid bacterium-containing oil / fat composition of the present invention include chocolate, glaze, whipped cream, sand cream, butter cream, filling and the like, and chocolate is particularly preferable. In the present specification, chocolate is not limited to the regulations (fair competition regulations regarding the labeling of chocolates) or the provisions of laws and regulations, and is mainly composed of raw materials derived from cocoa, and if necessary, sugars, dairy products, and other ingredients. Adds edible fats and oils, flavors, emulsifiers, etc. In the present specification, chocolate or chocolate confectionery is a chocolate-flavored chocolate confectionery such as dark chocolate, milk chocolate, white chocolate, colored chocolate based on white chocolate with a desired color, and chocolate confectionery containing these. Including all sweets.

Moreover, one embodiment of the present invention is a food containing the lactic acid bacterium-containing oil / fat composition of the present invention. Examples thereof include lactic acid bacterium-containing chocolate confectionery containing fruits, nuts, grains, gummies, candy and the like, baked confectionery containing lactic acid bacterium-containing oil and fat composition, snack confectionery, cookies, cakes, ice cream, beverages and the like.
Moreover, one embodiment of the present invention is a lactic acid bacterium-containing oil / fat composition characterized by containing a center material. In the present embodiment, the center material is coated with a lactic acid bacterium-containing oil / fat composition. As the center material, fruits, nuts, grains, gummies, candy and the like can be used.

Further, one embodiment of the present invention is a method for producing lactic acid bacterium-containing chocolate, which comprises adding lactic acid bacterium powder to a chocolate dough after tempering and before molding. is there.
In the method for producing chocolate containing lactic acid bacteria, when the lactic acid bacteria powder is added, the temperature of the chocolate dough is preferably 27 ° C. or higher and 31 ° C. or lower, more preferably 28 ° C. or higher and 30 ° C. or lower.

In the present embodiment, a chocolate having a high concentration of lactic acid bacteria is produced by mixing the powder of lactic acid bacteria with chocolate at a high concentration, and the chocolate containing a high concentration of lactic acid bacteria can be added to the chocolate dough after tempering and before molding. In this case, when chocolate containing a high concentration of lactic acid bacteria is added, the temperature of the chocolate dough before molding is preferably 27 ° C. or higher and 31 ° C. or lower, more preferably 28 ° C. or higher and 30 ° C. or lower. If the temperature is lower than 27 ° C, the chocolate dough will harden, and if the temperature exceeds 31 ° C, the crystal structure due to tempering will be broken and the quality cannot be maintained.
However, the method of the above embodiment is suitable for producing a molding type lactic acid bacterium-containing chocolate. As a further embodiment of the present invention, there is provided a method for producing lactic acid bacteria-containing chocolate that coats a center material. Since tempering is not performed in the production of center material-coated chocolate, the present embodiment is characterized in that lactic acid bacteria powder is added to a chocolate dough whose temperature has been adjusted to 35 ° C. or higher and 40 ° C. or lower, and the center material is coated with this. Provided is a method for producing chocolate containing lactic acid bacteria coated with a center material.

Example 1 Chocolate containing lactic acid bacteria (Labre bacteria)
1. 1. Preparation of Lactobacillus powder Lactobacillus powder is Lactobacillus brevis subsp. Coagulans was used. Labre bacteria were cultivated using a general lactic acid bacterium culture medium (MRS medium, etc.), and starch was added to a powder obtained by freeze-drying and crushing the bacterium collected by centrifugation or the like to obtain lactic acid bacterium powder. .. It is desirable that this lactic acid bacterium powder contains live lactic acid bacteria in an amount of 1 × 10 ⁵ cells / gram or more and 1 × 10 ¹³ cells / gram or less, and further, 1 × 10 ⁷ cells / gram or more 1 ×. It is preferable to contain 10 ¹³ pieces / gram or less, more preferably 1 × 10 ⁸ pieces / gram or more and 1 × 10 ¹³ pieces / gram or less.

2. Manufacture of chocolate containing lactic acid bacteria Generally, chocolate is made by selecting, separating, roasting, and grinding the raw material cocoa beans into cocoa mass, and cocoa mass and a part of sugar, powdered milk, vegetable oil, cocoa butter, and emulsifier. The chocolate dough is prepared by mixing with a raw material mixer, atomizing with a refiner, scouring (conching), and adding fragrance, lactic acid bacteria powder, and the remaining cocoa butter and emulsifier in the latter half of the scouring. In some cases, this chocolate dough is stored in a tank at about 45 to 50 ° C. for about 4 days on average. This storage is, so to speak, a waiting time caused by the convenience of operation of the production line. After that, the temperature is adjusted to about 28 to 30 ° C and tempered, and then the chocolate is filled in a molding mold, cooled and solidified, die-cut, and the obtained chocolate is packaged and inspected, and then shipped after aging. ..

Based on this production method, lactic acid bacterium-containing chocolate was produced by adding lactic acid bacterium powder by the conventional production method (adding lactic acid bacterium powder at the same timing as the addition of flavor or the like), production method 1 and production method 2. The outline of each manufacturing method is shown in FIG.

Conventional manufacturing method After refining, lactic acid bacteria powder was added to the chocolate dough that had reached 45 to 50 ° C, and after storage for 4 days, tempering and molding were performed.

Manufacturing method 1
Lactic acid bacteria powder was added to the chocolate dough (27 to 31 ° C.) that had been tempered.

Manufacturing method 2
A chocolate dough containing a high concentration of lactic acid bacteria was produced by dissolving the chocolate dough at 40 ° C. in advance and adding lactic acid bacteria powder to a high concentration. It is preferable that the chocolate containing a high concentration of lactic acid bacteria contains 1 × 10 ⁵ pieces / gram or more and 1 × 10 ¹² pieces / gram or less, and further 1 × 10 ⁷ pieces / gram. above 1 × ^{10 12} pieces / g or less, more preferably contained 1 × ^{10 12} pieces / g or less 1 × 10 ⁸ cells / g or more. The tempered chocolate dough was heated to 27 to 31 ° C., and chocolate containing a high concentration of lactic acid bacteria was added and mixed.

In any of the above-mentioned production methods, production was carried out according to the formulation of milk chocolate in Table 1.

Lactic acid bacteria-containing chocolate could be produced by any of the above production methods. When the production method 2 is used, it is possible to prevent lactic acid bacteria from scattering and being contaminated on the production line, so that the production method 2 is more preferable from the viewpoint of production control. Further, according to the method of the production method 2, the lactic acid bacteria can be uniformly mixed with the chocolate dough in a shorter time.
In addition to lactic acid bacteria powder, lactic acid bacteria-containing chocolate contains cocoa mass: 0 to 70% by weight, whole milk powder: 0 to 20% by weight, cocoa butter: 0 to 25% by weight, vegetable oil: 0 to 20% by weight, sugar. : Can be appropriately changed within the range of 0 to 45% by weight. In addition, auxiliary raw materials such as emulsifiers and fragrances can be added as appropriate, and the blending amounts thereof can be adjusted as appropriate. Further, dietary fiber: 0 to 15% by weight and oligosaccharide 0 to 10% by weight can be blended.

3. 3. Artificial gastric juice resistance test of lactic acid bacterium-containing chocolate Based on the production method 2, lactic acid bacterium-containing chocolate was prepared so that the lactic acid bacterium powder was 0.25% by weight. That is, according to Table 1, only the content ratio of lactic acid bacteria powder was changed. Using this as a sample, an artificial gastric juice resistance test was performed based on the following procedure.

(1) The MRS medium adjusted to a predetermined pH and added with 0.04% pepsin was used as artificial gastric juice (according to Non-Patent Document 1). The medium was 300 ml. The above artificial gastric juice was adjusted to pH 2.5 with dilute hydrochloric acid. Since the main component of gastric acid is hydrochloric acid, no other acid is used.

(2) Immerse 1 grain (2 grams) of chocolate in artificial gastric juice kept at 37 ° C, shake until the liquid level shakes lightly, and after 30 minutes, 1 hour and 2 hours, take 1 ml of artificial gastric juice and contain raw. The number of bacteria was measured. The method for measuring the viable cell count is the same as the time-dependent test for the viable lactic acid bacterium count. The maximum time is 2 hours because it usually takes about 2 hours for the digested material in the stomach to be completely transferred to the duodenum. As a control, a lactic acid bacterium-containing beverage containing Labre bacteria was also subjected to the same experiment by appropriately adjusting the addition amount so that the viable cell count at the time of addition in the artificial gastric juice was about the same as that of the lactic acid bacterium-containing chocolate.

(3) Results Since the same experiments were conducted three times on different dates and times, the results are shown in Table 2. The results of graphing these are shown in FIG. In the third experiment, two types of lactic acid bacteria-containing beverages containing Labre bacteria were used as comparative controls.

Surprisingly, from the results of the artificial gastric juice resistance test of lactic acid bacteria-containing chocolate, the lactic acid bacteria-containing chocolate produced by the manufacturing method 2 has a very high survival rate of lactic acid bacteria in the artificial gastric juice, and the survival rate is the control lactic acid bacteria. It turned out to be much more expensive than beverages.

4. Comparative test of resistance to artificial gastric fluid between chocolate containing lactic acid bacteria and powder of lactic acid bacteria Similar to the artificial gastric fluid resistance test of chocolate containing lactic acid bacteria, lactic acid bacteria-containing chocolate prepared so that the amount of powdered lactic acid bacteria was 0.25% by weight based on manufacturing method 2. Using the lactic acid bacteria powder itself, an artificial gastric fluid resistance test was performed in the same manner as the artificial gastric fluid resistance test for chocolate containing lactic acid bacteria.
However, as a control, lactic acid bacteria powder was used, and the number of viable bacteria at the time of addition in artificial gastric juice was appropriately adjusted to be about the same as that of chocolate containing lactic acid bacteria, and the same experiment was conducted.
The results are shown in Table 3. The result shown in the graph is shown in FIG.

In the artificial gastric juice resistance test of this example, the survival rate of lactic acid bacteria in the chocolate containing lactic acid bacteria of the present invention after 30 minutes, 1 hour and 2 hours, that is, the artificial gastric juice resistance is much higher than that of lactic acid bacteria powder. I found out.

5. Time-lapse test of the number of viable lactic acid bacteria in chocolate containing lactic acid bacteria Similar to the artificial gastric juice resistance test of chocolate containing lactic acid bacteria, lactic acid bacteria-containing chocolate containing 0.25% by weight of lactic acid bacteria powder is produced in chocolate based on the manufacturing method 2. did.

The number of viable lactic acid bacteria in the sample immediately after the production of this lactic acid bacterium-containing chocolate or stored at room temperature (about 18 ° C.) for 1 to 14 months was examined by the following method. As a control, a similar experiment was performed on soft candy containing lactic acid bacteria.

(1) One piece or about 1 g of a sample was taken, weighed precisely, put into 100 ml of a diluted solution, shaken vigorously, and uniformly suspended as a sample stock solution. 1 ml of the sample stock solution was added to 9 ml of the separately dispensed diluent to dilute the sample 10-fold. The same operation was repeated to prepare a sample solution.
(2) An appropriate amount of 1 ml or less of the sample solution was dispensed into two Petri dishes, and MRS agar medium kept at 50 ° C. was added and mixed to solidify.
(3) After solidification, anaerobic culture was carried out at 35 to 37 ° C. for 48 to 72 hours, the number of colonies that appeared was counted, and the average number of colonies was determined.
(4) The viable cell count (CFU) in 1 g of the sample was calculated from the following formula.
Viable cell count (CFU / g) = average number of colonies (CFU) x dilution ratio x sample stock solution (ml) / sample solution addition amount (ml) / sample collection amount (g)
(5) Results The experiment was conducted at room temperature (about 18 ° C). The results are shown in Table 4.

Table 5 shows the results of a time-dependent test on the number of lactic acid bacteria in the lactic acid bacteria-containing soft candy.

In soft candy, most of the lactic acid bacteria were killed one month after production, and there were almost no live bacteria two months later. It can be said that the residual rate is almost 0% both one month and two months after production. In comparison, chocolate containing lactic acid bacteria was able to maintain the viable state of lactic acid bacteria extremely stably. In addition, looking at the results of the sample 14 months after production at room temperature (about 18 ° C.), the stability of the lactic acid bacteria is good, so that the viable state of the lactic acid bacteria can be guaranteed for one year at room temperature.

6. Comparison of the Number of Lactic Acid Bacteria Contained in Lactic Acid Bacteria-Containing Chocolate Produced by Each Production Method The number of lactic acid bacteria per gram of lactic acid bacterium-containing chocolate produced by the conventional production methods, production methods 1 and 2, respectively, was measured. Table 6 shows the theoretical value of lactic acid bacteria calculated from the number of lactic acid bacteria contained in the added powder of lactic acid bacteria and the measured value.

As for the number of bacteria in the conventional manufacturing method, the number of lactic acid bacteria decreased due to the inactivation of the bacteria in the manufacturing process. Manufacturing method 1 and manufacturing method 2 are excellent manufacturing methods that increase the number of bacteria that reach the intestines alive because inactivation in the process can be prevented and the difference in the number of lactic acid bacteria is about 5 times that of the conventional manufacturing method. It can be said that. The viable number of lactic acid bacteria after being left at 45 ° C. for 2 days for chocolate containing a high concentration of lactic acid bacteria containing ^{about 1.4 × 10 9} pieces / gram of lactic acid bacteria powder produced in the production process of manufacturing method 2. , as a result of measurement by the same method, about 1.3 × 10 ⁹ cells / g, and almost no change. From this, it can be said that the decrease in the number of lactic acid bacteria in the conventional production method is not simply due to heat, and it is considered that long-term stirring during storage, tempering operation, etc. affect the decrease in the number of bacteria.

7. Artificial gastric juice resistance test of lactic acid bacteria-containing chocolate produced by each manufacturing method Using lactic acid bacteria-containing chocolate produced based on the conventional manufacturing methods, manufacturing methods 1 and 2, an artificial gastric juice resistance test is performed in the same procedure as the artificial gastric juice resistance test of lactic acid bacteria-containing chocolate. went.

Results Since the number of bacteria at the time of production differs depending on each production method, the number of bacteria at the time of production of each production method is set to 100%, and the results expressed as the residual rate are shown in Table 7.

From the above results, surprisingly, the lactic acid bacterium-containing chocolate produced by the production methods 1 and 2 was found to have a higher residual rate of lactic acid bacteria than the conventional production method in the artificial gastric juice resistance test. Therefore, it was found that the production methods 1 and 2 are excellent production methods also in terms of resistance to gastric acid.

As described above, from the results in this example, surprisingly, the method for producing lactic acid bacteria-containing chocolate according to the present invention is a very superior method for maintaining a high number of bacteria at the time of production, although it is a simple method. , It was found that it is a very excellent production method having higher resistance to gastric acid than lactic acid bacteria beverages and the like, and further having higher acid resistance than lactic acid bacteria-containing chocolate produced by a conventional production method. In addition, it was found that the lactic acid bacterium-containing chocolate produced by the production method is a chocolate having a higher effect of probiotics, and is a highly palatable and highly useful food for maintaining health.

Example 2 Chocolate containing lactic acid bacteria (lactic acid bacteria other than Labre bacteria)
1. 1. Production of chocolate containing lactic acid bacteria Based on the production method 2 with the formulation shown in Table 1, other lactic acid bacteria, namely Lactobacillus acidophilus, Lactobacillus gasseri, Enterococcus faecalis, and Biecalis. Change to Bifidobacterium longum or Leuconostoc mesenteroides so that the lactic acid bacteria powder is 0.5% by weight for Leuconostoc mesenteroides and 0.25% by weight for the other 4 types of bacteria. Lactobacillus-containing chocolate was produced in Japan.

2. Time-lapse test of viable lactic acid bacteria in chocolate containing lactic acid bacteria Similar to the time-lapse test of viable lactic acid bacteria in chocolate containing lactic acid bacteria in Example 1, the produced chocolate containing lactic acid bacteria was immediately after production of the chocolate containing lactic acid bacteria or at room temperature. The number of viable lactic acid bacteria in the sample stored at about 18 ° C. to about 25 ° C. for 1 to 3 months was examined. The results are shown in Table 8 below. It was found that the viable state can be maintained extremely stably for various bacteria 3 months after production in chocolate containing various bacteria.

3. 3. Artificial gastric juice resistance test of lactic acid bacteria-containing chocolate The produced lactic acid bacteria-containing chocolate was subjected to an artificial gastric juice resistance test of lactic acid bacteria-containing chocolate in the same manner as in Example 1. In addition, as a control, the same experiment was conducted for each bacterial powder. The results are shown in Table 9 below. It was found that the resistance of lactic acid bacteria to artificial gastric juice was much higher in chocolate containing various bacteria than the powder of lactic acid bacteria.

Example 3 Almond chocolate containing lactic acid bacteria 1. Production of Almond Chocolate Containing Lactic Acid Bacteria Labre bacterium was used as the lactic acid bacterium, and the labre bacterium powder was prepared in the same manner as in Example 1. The blending ratio is as shown in Table 10 below. As shown in FIG. 2, the outline of the production method was that the chocolate dough stored at 42 ° C. to 43 ° C. was adjusted to a temperature of 37 ° C. to 38 ° C., lactic acid bacteria powder was added, and the chocolate dough was used for almond chocolate hanging. In addition to lactic acid bacteria powder, sugar: 0 to 45% by weight, whole milk powder: 0 to 20% by weight, cocoa mass: 0 to 70% by weight, cocoa butter: 0 to 25% by weight, plants. Oils and fats: Can be appropriately changed within the range of 0 to 20% by weight. In addition, auxiliary raw materials such as emulsifiers and fragrances can be added as appropriate, and the blending amounts thereof can be adjusted as appropriate. Further, dietary fiber: 0 to 15% by weight and oligosaccharide 0 to 10% by weight can be blended. The type of center material is not limited to almonds and can be changed as appropriate, and the blending amount of the center material can also be adjusted as appropriate.

2. Time-lapse test of the number of viable lactic acid bacteria in the lactic acid bacteria-containing almond chocolate With respect to the produced lactic acid bacteria-containing almond chocolate, immediately after the production of the lactic acid bacteria-containing almond chocolate, as in the time-lapse test of the number of viable lactic acid bacteria in the lactic acid bacteria-containing chocolate of Example 1. Alternatively, the number of viable lactic acid bacteria in the sample stored at room temperature (about 18 ° C.) for 3 to 7 months was examined. The results are shown in Table 11 below, and it was found that the viable state can be maintained extremely stably.

3. 3. Artificial gastric juice resistance test of lactic acid bacteria-containing almond chocolate The produced lactic acid bacteria-containing almond chocolate was subjected to an artificial gastric juice resistance test of lactic acid bacteria-containing almond chocolate in the same manner as in Example 1. The results are shown in Table 12 below. As a control, the same experiment was conducted for lactic acid bacteria powder. It was found that the resistance of lactic acid bacteria to artificial gastric juice was much higher in almond chocolate containing lactic acid bacteria than that of lactic acid bacteria powder.

Example 4 Sugarless chocolate containing lactic acid bacteria In this example, sugarless chocolate containing lactic acid bacteria was produced by the formulation shown in Table 13 below based on the production method 2.

The lactic acid bacterium-containing sugarless chocolate also showed almost the same time stability and artificial gastric juice resistance as in Example 1 in the lactic acid bacterium-containing time-varying test and the artificial gastric juice resistance test.
In addition to lactic acid bacteria powder, sugar alcohol: 0 to 50% by weight, cocoa mass: 0 to 70% by weight, milk powder: 0 to 25% by weight, vegetable oil: 0 to 25% by weight, Cocoa butter: Can be changed as appropriate within the range of 0 to 25% by weight. In addition, auxiliary raw materials such as high-sweetness sweeteners, emulsifiers, and flavors can be added as appropriate, and the blending amounts thereof can be adjusted as appropriate. Further, dietary fiber: 0 to 15% by weight and oligosaccharide: 0 to 10% by weight can be blended. The oligosaccharides that can be used include, but are not limited to, fructooligosaccharides, galactooligosaccharides, isomaltooligosaccharides, and soybean oligosaccharides.

Example 5 Lactic acid bacteria-containing fat and oil composition 1. Production of Lactic Acid Bacteria-Containing Oil / Fat Composition A lactic acid bacterium-containing oil / fat composition was produced using Labre bacterium powder prepared in the same manner as in Example 1. The blending amount is as shown in Table 14 below. The outline of the manufacturing method is shown in FIG. This lactic acid bacterium-containing oil / fat composition is solid near room temperature (25 ° C.), but becomes liquid near human body temperature (35 ° C. to 40 ° C.) and has physical characteristics similar to those of hand cream.
In addition to the lactic acid bacteria powder, the composition of the lactic acid bacterium-containing fat and oil composition includes sugar: 0 to 80% by weight, vegetable oil and fat: 20 to 99% by weight, whole milk powder: 0 to 40% by weight, and lactose: 0 to 20% by weight. It can be changed as appropriate within the range. In addition, auxiliary raw materials such as high-sweetness sweeteners, emulsifiers, and flavors can be added as appropriate, and the blending amounts thereof can be adjusted as appropriate. Further, dietary fiber: 0 to 15% by weight and oligosaccharide: 0 to 10% by weight can be blended. This lactic acid bacterium-containing oil / fat composition is white chocolate or a composition similar to white chocolate, and can be used as a coating or topping for foods such as baked goods and chocolate, and as a material for decoration. Not limited to.

2. Time-lapse test of the number of viable lactic acid bacteria in the lactic acid bacteria-containing fat and oil composition With respect to the produced lactic acid bacteria-containing fat and oil composition, the time-dependent test of the number of viable lactic acid bacteria in the lactic acid bacteria-containing chocolate of Example 1 was carried out. The number of viable lactic acid bacteria in a sample immediately after production or stored at room temperature (about 25 ° C.) for 1 to 2 months was examined. The results are shown in Table 15 below, and it was found that the viable state can be maintained extremely stably.

3. 3. Artificial gastric juice resistance test of lactic acid bacterium-containing fat and oil composition The artificial gastric juice resistance test of the lactic acid bacterium-containing fat and oil composition was performed on the produced lactic acid bacterium-containing fat and oil composition in the same manner as in Example 1. The results are shown in Table 16 below. As a control, the same experiment was conducted for lactic acid bacteria powder. It was found that the resistance of lactic acid bacteria to artificial gastric juice in the lactic acid bacteria-containing oil / fat composition was much higher than that of lactic acid bacteria powder.

This application claims priority from Japanese Patent Application No. 2015-111706 filed on June 1, 2015, and the contents are cited as part of this application.

Claims (4)

A chocolate containing a high concentration of lactic acid bacteria containing 1 × 10 ⁵ pieces / g or more and 1 × 10 ¹² pieces / g or less of lactic acid bacteria was produced by mixing lactic acid bacteria powder with chocolate at a high concentration.
Add chocolate with high concentration of lactic acid bacteria to the chocolate dough after tempering and before molding.
A method for producing chocolate containing lactic acid bacteria, wherein the temperature of the chocolate dough after tempering and before molding is 27 ° C. or higher and 31 ° C. or lower when chocolate containing a high concentration of lactic acid bacteria is added. The method for producing lactic acid bacteria-containing chocolate according to claim 1, wherein when the chocolate containing a high concentration of lactic acid bacteria is added, the temperature of the chocolate dough after tempering and before molding is 28 ° C. or higher and 30 ° C. or lower. The content of the lactic acid bacteria is 1 × 10 ⁷ Pieces / g or more 1 x 10 ¹² The method for producing lactic acid bacteria-containing chocolate according to claim 1 or 2, wherein the number of pieces / g or less. The content of the lactic acid bacteria is 1 × 10 ⁸ Pieces / g or more 1 x 10 ¹² The method for producing chocolate containing lactic acid bacteria according to claim 3, wherein the number of pieces / g or less.

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